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米蛾体内WO噬菌体和Wolbachia的感染检测及其在组织中的分布

发表时间:2016-05-15  浏览量:1721  下载量:599
全部作者: 丛 斌,张明珠,李帅强,张统书,董 辉
作者单位: 沈阳农业大学植物保护学院
摘 要: 目的:Wolbachia各品系中存在广泛的WO噬菌体侵染现象,推测其很可能参与了Wolbachia遗传变异和Wolbachia对寄主生殖调控的过程。本实验对米蛾(Corcyra cephalonica)成虫进行了Wolbachia和WO噬菌体的感染检测,并探究两者在米蛾各组织中的分布情况,为以后深入研究“WO噬菌体-Wolbachia-寄主”三者的关系奠定基础。方法:利用SDS蛋白酶K法提取米蛾成虫总DNA及各组织DNA,通过Wolbachia表面蛋白基因(wsp)和WO噬菌体衣壳蛋白基因(orf 7)特异性引物检测米蛾体内的Wolbachia和WO噬菌体感染情况,并明确两者在头、胸、腹、足、翅等组织中的分布。结果:wsp基因特异性引物只在米蛾中扩增到一条602 bp的序列,orf 7基因特异性引物获得两条长度分别为409 bp和424 bp的序列;在米蛾头、胸、腹、足、翅等组织中两特异性引物均能成功扩增。结论:米蛾头、胸、腹、足、翅等组织中均存在Wolbachia和WO噬菌体感染。米蛾只感染一种Wolbachia,分子系统发育树分析显示其属于B超组Pip亚组;Blasting比对分析显示,米蛾体内Wolbachia受到两类WO噬菌体的侵染,将其命名为WOcep1(409 bp)和WOcep2(424 bp).
关 键 词: 农业昆虫学;米蛾;Wolbachia;WO噬菌体;wsp;orf 7
Title: Detection of phage WO and Wolbachia in Corcyra cephalonica and both distribution in different organs
Author: CONG Bin, ZHANG Mingzhu, LI Shuaiqiang, ZHANG Tongshu, DONG Hui
Organization: College of Plant Protection, Shenyang Agricultural University
Abstract: Objective: Many Wolbachia strains were found infected by phage WO. It is postulated that phage WO involves in the control of genetic variation of Wolbachia and the regulation of reproduction of the hosts. To further understand the interactions between phage WO, Wolbachia and the hosts, laboratory tests were conducted to detect the presence of phage WO and Wolbachia in different organs of Corcyra cephalonica. Methods: Total DNAs of different organs and the adult C. cephalonica were extracted using SDS-proteinase K method; PCR technique was used to detect the infection of phage WO and Wolbachia with primers of Wolbachia surface protein gene (wsp) and putative phage capsid protein gene (orf 7). Furthermore, the distributions of phage WO and Wolbachia in head, thorax, abdomen, leg and ala were also analyzed. Results: For the whole body of C. cephalonica, wsp sequence was specifically amplified as 602 bp; the orf 7 primer amplified two sequences (409 bp and 424 bp) in total DNA of the whole body. In addition, PCR with these two primers also successfully amplified their target fragments in different organs. Conclusion: The head, thorax, abdomen, leg and ala of C. cephalonica were all infected by Wolbachia and phage WO. The phylogenetic tree constructed with wsp gene showed that all Wolbachia in C. cephalonica belong to Pip subgroup; Blasting search analysis showed that Wolbachia in C. cephalonica were infected by two kinds of phage WO, which were WOcep1 (409 bp) and WOcep2 (424 bp), respectively.
Key words: agricultural entomology; Corcyra cephalonica; Wolbachia; phage WO; wsp; orf 7
发表期数: 2016年5月第9期
引用格式: 丛 斌,张明珠,李帅强,等. 米蛾体内WO噬菌体和Wolbachia的感染检测及其在组织中的分布[J]. 中国科技论文在线精品论文,2016,9(9):933-941.
 
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